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Publication
Recombinant SARS-CoV-2 envelope protein traffics to the trans-Golgi network
following amphipol-mediated delivery into human cells.
Authors Hutchison JM, Capone R, Luu DD, Shah KH, Hadziselimovic A, Van Horn WD, Sanders
CR
Submitted By Submitted Externally on 12/3/2021
Status Published
Journal The Journal of biological chemistry
Year 2021
Date Published 8/1/2021
Volume : Pages 297 : 100940
PubMed Reference 34237302
Abstract The severe acute respiratory syndrome coronavirus 2 envelope protein (S2-E) is a
conserved membrane protein that is important for coronavirus (CoV) assembly and
budding. Here, we describe the recombinant expression and purification of S2-E
in amphipol-class amphipathic polymer solutions, which solubilize and stabilize
membrane proteins, but do not disrupt membranes. We found that amphipol delivery
of S2-E to preformed planar bilayers results in spontaneous membrane integration
and formation of viroporin cation channels. Amphipol delivery of the S2-E
protein to human cells results in plasma membrane integration, followed by
retrograde trafficking to the trans-Golgi network and accumulation in swollen
perinuclear lysosomal-associated membrane protein 1-positive vesicles, likely
lysosomes. CoV envelope proteins have previously been proposed to manipulate the
luminal pH of the trans-Golgi network, which serves as an accumulation station
for progeny CoV particles prior to cellular egress via lysosomes. Delivery of
S2-E to cells will enable chemical biological approaches for future studies of
severe acute respiratory syndrome coronavirus 2 pathogenesis and possibly even
development of "Trojan horse" antiviral therapies. Finally, this work also
establishes a paradigm for amphipol-mediated delivery of membrane proteins to
cells.




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